Difference between revisions of "Team:Aix-Marseille/Composite Part"
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<!-- Pour chaque part il serait bien d'expliquer d'ou vien la sequence (les parts utilisé pour le faire), ce que c'est censé faire (pourquoi on la construit), comment on a testé ce que ca fait essentiellement copier coller du registry --> | <!-- Pour chaque part il serait bien d'expliquer d'ou vien la sequence (les parts utilisé pour le faire), ce que c'est censé faire (pourquoi on la construit), comment on a testé ce que ca fait essentiellement copier coller du registry --> | ||
| − | =BBa_K2718006= | + | ==BBa_K2718006== |
Methionine gamma-lyase (MGL) catalyzes the conversion of l-methionine to methanethiol, the oxidation of methanethiol then produces first DMDS (Dimethyl disulfide) and then DMTS ( Dimethyl trisulfide). The composite part [http://parts.igem.org/Part:BBa_K2718006 BBa_K2718006] is made from [http://parts.igem.org/Part:BBa_K2718005 BBa_K2718005], an RFC 10 compatible part that we derived from [http://parts.igem.org/Part:BBa_K1493300 BBa_K1493300] using a PCR to add a C-terminal histidine tag, preceded by the [http://parts.igem.org/wiki/index.php?title=Part:BBa_R0011 BBa_R0011], an IPTG inducible promoter part. | Methionine gamma-lyase (MGL) catalyzes the conversion of l-methionine to methanethiol, the oxidation of methanethiol then produces first DMDS (Dimethyl disulfide) and then DMTS ( Dimethyl trisulfide). The composite part [http://parts.igem.org/Part:BBa_K2718006 BBa_K2718006] is made from [http://parts.igem.org/Part:BBa_K2718005 BBa_K2718005], an RFC 10 compatible part that we derived from [http://parts.igem.org/Part:BBa_K1493300 BBa_K1493300] using a PCR to add a C-terminal histidine tag, preceded by the [http://parts.igem.org/wiki/index.php?title=Part:BBa_R0011 BBa_R0011], an IPTG inducible promoter part. | ||
| − | To test the activity of [http://parts.igem.org/Part:BBa_K2718006 BBa_K2718006] we produced and purified the his-tagged protein and used DTNB to measure the enzyme activity. DTNB reacts with the released thiol group [https://www.dovepress.com/activities-of-methionine-gamma-lyase-in-the-acidophilic-archaeon-ldquo-peer-reviewed-article-RRB]. | + | To test the activity of [http://parts.igem.org/Part:BBa_K2718006 BBa_K2718006] we produced and purified the his-tagged protein and used DTNB to measure the enzyme activity. DTNB reacts with the released thiol group <ref>[https://www.dovepress.com/activities-of-methionine-gamma-lyase-in-the-acidophilic-archaeon-ldquo-peer-reviewed-article-RRB]</ref>. |
| − | =BBa_K2718010= | + | ==BBa_K2718010== |
| − | =BBa_K2718011= | + | ==BBa_K2718011== |
| − | =BBa_K2718022= | + | ==BBa_K2718022== |
Our part [http://parts.igem.org/Part:BBa_K2718022 BBa_K2718022] is made by fusing the promoter RBS in part | Our part [http://parts.igem.org/Part:BBa_K2718022 BBa_K2718022] is made by fusing the promoter RBS in part | ||
[http://parts.igem.org/Part:BBa_J04500 BBa_J04500] to our chitinase part with a C-terminal oligo-histidine tag [http://parts.igem.org/Part:BBa_K2718021 BBa_K2718021]. | [http://parts.igem.org/Part:BBa_J04500 BBa_J04500] to our chitinase part with a C-terminal oligo-histidine tag [http://parts.igem.org/Part:BBa_K2718021 BBa_K2718021]. | ||
| − | We tested the enzyme activity using Schales' method[https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3975300/pdf/1754-6834-7-37.pdf] we also tested production of the protein and used the his-tag to purify the protein. | + | We tested the enzyme activity using Schales' method<ref>[https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3975300/pdf/1754-6834-7-37.pdf]</ref> we also tested production of the protein and used the his-tag to purify the protein. |
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| + | ==References== | ||
| + | <references /> | ||
Revision as of 18:00, 17 October 2018
Composite Parts
BBa_K2718006
Methionine gamma-lyase (MGL) catalyzes the conversion of l-methionine to methanethiol, the oxidation of methanethiol then produces first DMDS (Dimethyl disulfide) and then DMTS ( Dimethyl trisulfide). The composite part BBa_K2718006 is made from BBa_K2718005, an RFC 10 compatible part that we derived from BBa_K1493300 using a PCR to add a C-terminal histidine tag, preceded by the BBa_R0011, an IPTG inducible promoter part. To test the activity of BBa_K2718006 we produced and purified the his-tagged protein and used DTNB to measure the enzyme activity. DTNB reacts with the released thiol group [1].
BBa_K2718010
BBa_K2718011
BBa_K2718022
Our part BBa_K2718022 is made by fusing the promoter RBS in part BBa_J04500 to our chitinase part with a C-terminal oligo-histidine tag BBa_K2718021. We tested the enzyme activity using Schales' method[2] we also tested production of the protein and used the his-tag to purify the protein.